AOAC-RI ERP Book MICRO Sept 2016.pdf
1568 B ird et al . : J ournal of aoaC i nternational V ol . 97, n o . 6, 2014
Table 2014.01C. Sample matrix and enrichment scheme a
Sample size, g
Enrichment broth volume, mL
Enrichment time, h
Secondary enrichment time, h
Sample matrix
Raw ground beef (80% lean)
25 25
225 225
18–24 18–24
8–24 8–24
Raw ground chicken
Raw ground pork
25
225
18–24
8–24
Frozen uncooked shrimp
25
225
18–24
8–24
Fresh bunched spinach
25
225 225
18–24 18–24
24
Stainless steel; environmental sponges
1 Sponge (4 × 4 in.)
Pasteurized liquid whole egg
100
900
18–24
Cooked breaded chicken
325
2925
18–24
Dry dog food
375
3375
18–24
a AOAC RI Certificate No. 061301.
( b ) Follow all instructions carefully. Failure to do so may lead to inaccurate results. ( c ) After use, the enrichment medium and the 3M Petrifilm SALX Plates and 3M Petrifilm SALX Confirmation Disks can potentially contain pathogenic materials. When testing is complete, follow current industry standards for the disposal of contaminated waste. Consult the Material Safety Data Sheet for additional information and local regulations for disposal. D. Sample Enrichment ( 1 ) Prewarm 3M Salmonella Enrichment Base with 3M Salmonella Enrichment Supplement (50 mg/L) to 41.5 ± 1°C. ( 2 ) Aseptically combine the enrichment medium and sample following Table 2014.01C . For all meat and highly particulate samples, the use of filter bags is recommended. Homogenize thoroughly for 2 min and incubate at 41.5 ± 1°C for 18–24h. ( a ) Foods with high microbial backgrounds (≥10 4 CFU/g).— Transfer 0.1 mL of the primary enrichment into 10.0 mL R-V R10 broth. Incubate for 8–24 h at 41.5 ± 1°C. ( b ) Foods with low microbial backgrounds (<10 4 CFU/g).— Proceed to 3M Petrifilm SALX Plate preparation as described in E . E. Preparation of the 3M Petrifilm Salmonella Express Plates ( 1 ) Place the 3M Petrifilm SALX Plate on a flat, level surface. ( 2 ) Use prescribed diluents to hydrate the 3M Petrifilm SALX Plates: Butterfield’s Phosphate Diluent, distilled water, or reverse osmosis water. ( 3 ) Lift the top film and with the pipet perpendicular dispense 2.0±0.1 mL sterile diluent onto the center of bottom film. Do not close the top film before dispensing the entire 2.0 mL volume. ( 4 ) Gently roll down the top film onto the diluent to prevent trapping air bubbles. ( 5 ) Place the 3M Petrifilm Flat Spreader on the center of the plate. Press gently on the center of the spreader to distribute the diluent evenly. Spread the diluent over the entire 3M Petrifilm SALX Plate growth area before the gel is formed. Do not slide the spreader across the film.
hydrated 3M Petrifilm SALX Plates, and 3M Petrifilm SALX Plates after incubation. ( q ) Refrigerator.— Capable of maintaining 2–8°C for storing unopened 3M Petrifilm SALX Plates and 3M Petrifilm SALX Confirmation Disk. C. General Instructions ( a ) Store 3M Petrifilm SALX Plates and 3M Petrifilm SALX Confirmation Disks at 2–8°C. After opening the 3M Petrifilm SALX Plate pouches, seal the pouch and store at ambient temperature, less than 60% relative humidity (RH). Hydrated 3M Petrifilm SALX Plates can be stored up to 7 days at 2–8°C. Post-incubation 3M Petrifilm SALX Plates can be stored at –10 to –20°C for up to 3 days. Hydrate the 3M Petrifilm SALX Plates with 2.0 ± 0.1 mL sterile diluent. Do not allow the top film to close before dispensing the entire 2.0 mL volume. Gently roll down the top film onto the diluent to prevent trapping air bubbles. Place the 3MPetrifilm Flat Spreader on the center of the plate. Press gently on the center of the spreader to distribute the diluent evenly. Spread the diluent over the entire 3M Petrifilm SALX Plate. Remove the spreader and leave the 3M Petrifilm SALX Plate undisturbed for 1 min. Prior to use, place the plates on a flat surface for 1 h at room temperature (20–25°C/<60% RH) and protected from light to allow the gel to form. Hydrated plates can be stored at room temperature (20–25°C/<60% RH) protected from light for up to 8 h before use.
Figure 2014.01. Streaking pattern on the 3M Petrifilm SALX Plate.
Figure 2014.01. Streaking pattern on the 3M Petrifilm Salmonella Express Plate.
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